Team:Tsinghua/Project

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''Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
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[[Image:project.png|center|frameless|Overall Project|500px]]
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!align="center"|[[Team:Tsinghua|Home]]
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!align="center"|[[Team:Tsinghua/Team|The Team]]
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!align="center"|[[Team:Tsinghua/Brainstorming|Brainstorming]]
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!align="center"|[[Team:Tsinghua/Project|The Project]]
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(''Or you can choose different headings.  But you must have a team page, a project page, and a notebook page.'')
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=Overall Project=
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Our aim is to construct a targeted gene therapy vector with high cellular specificity, considerable capacity and the potential for mass production and universal modification. Analogizing the characteristics of bacteriophage lambda and adenovirus, we genomically engineered the fiber protein of adenovirus with the pC of bacteriophage lambda, with the knob region modified by cell-specific peptides generated by phage display (called targeted bioBrick). After inducing the vector genome (generated by bottom-up or top-down approach) into BL21 DE3 E.coli strain, we applied a co-transformed therapeutic DNA (namely a cosmid with a capacity of 40-50 kb) for mass production of our targeted gene therapy vectors containing the desired genes to be delivered. With the targeted bioBrick mediating the attachment and RGD domain mediating the internalization of the targeted vector, we are able to accomplish the targeted gene therapy.
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== '''Overall project''' ==
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==ModuleI: Synthesis of GenSniper Viroin==
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Using two synthetic biological strageties (bottom-up and top-down), we will construct a genome of our desired gene therapy vector, which consists of the structural proteins for bacteriophage lambda head assembly and lysis genes (and targeted bioBrick later). Our aim in this module is to generate the viroin of our gene therapy vector (namely the head structure of bacteriophage lambda).
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Your abstract
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==ModuleII: Therapeutic DNA==
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In this module, we plan to construct a cosmid (a molecular cloning with a cos site) encoding the conceptual therapeutic genes, because the structural proteins of bacteriophage lambda will only package the circular DNA with cos site and 40-50 kb in size. Here we will also identify the function of this module through the expression of RFP.
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==ModuleIII: Targeted BioBrick==
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This module is the modification of bacteriophage lambda protein C by fiber shaft at its N termini. When constructing this targeted bioBrick, we will apply PCR to add the targeted peptide (in wetlab we just use TAT peptide) and RGD sequence into the fusion protein, forming a peptide-fiber shaft-RGD-C reading frame.
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{{Tsinghua/Pic}}
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== Background ==
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=== Gene Therapy ===
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Gene therapy is the insertion of genes into an individual's cells and tissues to treat a disease, such as a hereditary disease in which a deleterious mutant allele is replaced with a functional one[1]. Although the technology is still in its infancy, it is one of the most promising and active research fields in medicine[1,2]. Antisense therapy is not strictly a form of gene therapy, but is a genetically-mediated therapy and is often considered together with other methods[1].
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=== Vectors in Gene Therapy ===
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Despite substantial progress, a number of key technical issues need to be resolved before gene therapy can be safely and effectively applied in the clinic, and an ideal gene delivery vector is one of the bottlenecks in gene therapy application[].
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Generally, vectors applied in gene therapy can be classified into viral or non-viral.
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==== Viral Methods ====
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All viruses bind to their hosts and introduce their genetic material into the host cell as part of their replication cycle. This genetic material contains basic 'instructions' of how to produce more copies of these viruses, hijacking the body's normal production machinery to serve the needs of the virus[1].
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Adenovirus.
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Adeno-associated virus.
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Retroviruses.
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==== Non-Viral Methods ====
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Non-viral methods present certain advantages over viral methods, with simple large scale production and low host immunogenicity being just two. Previously, low levels of transfection and expression of the gene held non-viral methods at a disadvantage; however, recent advances in vector technology have yielded molecules and techniques with transfection efficiencies similar to those of viruses[1].
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Naked DNA.
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Oligonucleotides.
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Lipoplexes and polyplexes.
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=== Frontiers in Gene Therapy Research ===
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==== Induced Pluripotent Stem Cell (iPS) and Gene Delivery System====
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==== Therapeutic microRNA Delivery====
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Therapeutic strategies based on modulation of microRNA (miRNA) activity hold great promise due to the ability of these small RNAs to potently influence cellular behavior[3].
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==== Cancer Gene Therapy====
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As a primary threat of human health, cancer causes about 13% of all human deaths[4]. According to the American Cancer Society, 7.6 million people died from cancer in the world during 2007[5]. Current treatments often have far reaching negative side effects[6]. The systemic toxicity of chemotherapy regimens still often result in acute and delayed nausea, mouth ulcerations and mild cognitive impairments[7].
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== Project Details==
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=== Introduction ===
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=== Basic Idea ===
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=== The Experiments ===
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==== Project 1 ====
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==== Project 2 ====
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==== Project 1 and Project 2 ====
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== Results ==
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==== Project 1 ====
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==== Project 2 ====
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== References ==
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[1] http://en.wikipedia.org/wiki/Gene_therapy
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[2] SM Selkirk. Gene therapy in clinical medicine. ''Postgraduate Medical Journal''.2004;80:560-570; doi:10.1136/pgmj.2003.017764
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[3] Janaiah Kota, Raghu R. Chivukula, Kathryn A. O’Donnell, Erik A. Wentzel, Chrystal L. Montgomery, Hun-Way Hwang, Tsung-Cheng Chang, Perumal Vivekanandan, Michael Torbenson, K. Reed Clark, Jerry R. Mendell,and Joshua T. Mendell. Therapeutic microRNA Delivery Suppresses Tumorigenesis in a Murine Liver Cancer Model. ''Cell''. 2009, 137. 1005–1017.
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[4] WHO (February 2006). "Cancer". World Health Organization. http://www.who.int/mediacentre/factsheets/fs297/en/. Retrieved on 2007-06-25.
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[5] American Cancer Society (December 2007). "Report sees 7.6 million global 2007 cancer deaths". Reuters. http://www.reuters.com/article/healthNews/idUSN1633064920071217. Retrieved on 2008-08-07.
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[6] Peter Sinnaeve, Olivier Varenne, Désiré Collen, and Stefan Janssens. Gene therapy in the cardiovascular system: an update. ''Cardiovasc Res'', Dec 1999; 44: 498 - 506.
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[7] Chemotherapy and you: A guide to self-help during cancer treatment. National Institutes of Health Web site. Available at: http://www.cancer.gov/PDF/b21d0a74-b477-41ec-bdc0-a60bbe527786/chemoandyou.pdf. Accessed July 31, 2006.
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Latest revision as of 00:05, 22 October 2009

Home Team Project Modeling Parts ELSI

Home Background Brainstorming Design Experiment Results Conclusion Protocol


Overall Project


Contents

Overall Project

Our aim is to construct a targeted gene therapy vector with high cellular specificity, considerable capacity and the potential for mass production and universal modification. Analogizing the characteristics of bacteriophage lambda and adenovirus, we genomically engineered the fiber protein of adenovirus with the pC of bacteriophage lambda, with the knob region modified by cell-specific peptides generated by phage display (called targeted bioBrick). After inducing the vector genome (generated by bottom-up or top-down approach) into BL21 DE3 E.coli strain, we applied a co-transformed therapeutic DNA (namely a cosmid with a capacity of 40-50 kb) for mass production of our targeted gene therapy vectors containing the desired genes to be delivered. With the targeted bioBrick mediating the attachment and RGD domain mediating the internalization of the targeted vector, we are able to accomplish the targeted gene therapy.

ModuleI: Synthesis of GenSniper Viroin

Using two synthetic biological strageties (bottom-up and top-down), we will construct a genome of our desired gene therapy vector, which consists of the structural proteins for bacteriophage lambda head assembly and lysis genes (and targeted bioBrick later). Our aim in this module is to generate the viroin of our gene therapy vector (namely the head structure of bacteriophage lambda).

ModuleII: Therapeutic DNA

In this module, we plan to construct a cosmid (a molecular cloning with a cos site) encoding the conceptual therapeutic genes, because the structural proteins of bacteriophage lambda will only package the circular DNA with cos site and 40-50 kb in size. Here we will also identify the function of this module through the expression of RFP.

ModuleIII: Targeted BioBrick

This module is the modification of bacteriophage lambda protein C by fiber shaft at its N termini. When constructing this targeted bioBrick, we will apply PCR to add the targeted peptide (in wetlab we just use TAT peptide) and RGD sequence into the fusion protein, forming a peptide-fiber shaft-RGD-C reading frame.


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